This project will focus on expanding the therapeutic range of HSV-1 vectors for treatment of brain tumors and on[unreadable] creating zones of resistance to tumor growth in the normal brain by developing novel vectors and delivery modalities.[unreadable] The current proposal plans to increase the therapeutic capacity of the oncolytic HSV-1 vector, MGH2 in two ways: by[unreadable] enhancing infection of glioblastoma (GBM) cells through targeting surface antigens by virions; and by combining[unreadable] oncolytic vectors with amplifying/integrating HSV/AAV amplicon vectors to increase and sustain therapeutic transgene[unreadable] levels. We will also attempt to create a zone of resistance to tumor growth in normal brain via AAV vectors and[unreadable] neuroprecursor cells (MFCs) expressing secretable therapeutic proteins. NPCs and chimeric immune receptor (CIR)[unreadable] lymphocytes will be used to target invasive tumor cells. All studies will be carried out in culture and in intracranial[unreadable] human glioma models in nude mice using bioluminescence imaging reporters to track vector and protein delivery, cell[unreadable] fate and tumor growth. An immunocompetent syngeneic rat GBM model will also be used in evaluating CIR[unreadable] lymphocytes. Aim 1 will explore selective infection of glioblastoma cells expressing the mutant EGF receptor,[unreadable] EGFRvIII, by modification of the glycoprotein C envelope protein of HSV-1 virions for presentation of antibodies to[unreadable] this receptor. Aim 2 will evaluate the ability of CIR lymphocytes to target to EGFRvIII and VEGFR receptors in tumor[unreadable] foci. In Aim 3, AAV vectors and genetically modified NPCs will be used to deliver secretable forms of the tumorspecific[unreadable] apoptotic protein, TRAIL, and the anti-angiogenic factor, Flkl to normal brain to create a region of resistance[unreadable] to tumor growth. Aim 4 will employ a dual gene delivery system combining hybrid amplicon vectors incorporating p5[unreadable] and ITR DNA elements from AAV, and Rep 78/68 proteins as fusions with the HSV-1 virion tegument protein, VP16.[unreadable] Co-infection with oncolytic HSV-1 or a subset of HSV-1 genes will be evaluated for replicative amplification and[unreadable] genomic integration of amplicon-encoded transgenes. This project will use expertise on oncolytic virus (Project 1,[unreadable] Chiocca), in vivo imaging (Project 3, Weissleder), biostatistics (Core A, Finkelstein), HSV vector stocks (Core B,[unreadable] Krisky/Glorioso), and human GBM cells and neuropathology (Core C, Louis). These studies are designed to[unreadable] complement and increase the therapeutic impact of the HSV-1 oncolytic vector strategy for cancer treatment and to[unreadable] explore the possibility of creating regions of the brain resistant to tumor growth by sustained release of therapeutic[unreadable] proteins, using means deemed compatible with human trials.[unreadable]